Genotype evaluation,muscle histopathology and ultrastructure in a Duchenne muscular dystrophy animal model

AIM: To evaluate the genotype , muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.     

Selection of soybean elite cultivars based on phenotypic and genomic characters related to lodging tolerance

Soybean lodging can result in serious yield reduction. Detecting the quantitative trait loci (QTL) associated with lodging tolerance for their further application in marker‐assisted selection (MAS) has the potential to enhance soybean breeding efficiency. In this study, a genome‐wide association analysis (GWAS) was performed to identify soybean accessions that could potentially be used to produce lodging‐tolerant varieties, based on the comprehensive evaluation of lodging scores (LS) obtained for the parental cultivar “Tokachi nagaha” and its 137 derived cultivars. Results showed that genotype, environment and genotype × environment interaction significantly influenced LS. Of the 31 significant SNPs identified, 22 were consistently detected in two or more environments and 27 SNPs were located in or close to agronomically important QTL mapped by linkage analysis. Best linear unbiased predictors (BLUPs) of LS tend to decrease with the elite alleles contained by accessions increasing. Some excellent accessions, with lower BLUPs and D_i (stability coefficients) values and more elite alleles, were selected. This study contributed to understand the genetic mechanism of lodging, providing genetic and phenotypic information for MAS.     

不同饲养方式对肉鸡肌纤维组织学特性及肌肉嫩度影响的研究

[目的]研究了不同饲养方式对河田鸡肌纤维组织学特性及肌肉嫩度的影响,为肉鸡品质调控提供理论依据。[方法]研究户外散养、室内平养、室内笼养3种饲养方式对不同日龄河田鸡腿肌和胸肌肌纤维组织学特性的影响及其与肌肉嫩度的关系。[结果]结果表明,户外散养鸡的胸肌和腿肌肌纤维直径和肌肉剪切力显著大于笼养鸡（P〈0．05）,而肌纤维密度显著低于笼养鸡（P〈0．05）。随着日龄的增加,肌纤维的直径和肌肉剪切力显著增加,而肌纤维密度显著下降。公鸡的肌纤维直径和剪切力大于母鸡。相关性分析表明,胸肌肌肉剪切力与肌纤维直径呈显著正相关（P〈0．05）,而与肌纤维密度呈负相关（P〈0．05）;腿肌肌肉剪切力与肌纤维直径和密度关系不显著（P〉0．05）。[结论]饲养方式对肉鸡肌纤维的组织学特性有显著影响,户外散养促进肉鸡肌纤维发育,相应增加了肌肉剪切力。     

肌肉苹果酸酶活性与猪肉品质关系的研究

[目的]探讨肌肉苹果酸酶活性与猪肌肉品质的关系,为提高猪肌肉品质提供理论依据。[方法]选取3个不同品种的猪（鄂西黑猪、长野鄂杂交猪和长大二元杂交猪）,比较了肌肉苹果酸酶活性以及肉质性状之间的差异。[结果]结果表明,鄂西黑猪的肌肉苹果酸酶活性和肌内脂肪显著高于长野鄂（三元）杂交猪和长大二元杂交猪（P〈0.05）;长大二元杂交猪的肌肉pH2值、滴水损失和剪切力显著高于鄂西黑猪（P〈0.05）。[结论]猪肌肉苹果酸酶活性对猪肉品质存在一定的影响。其中,肌肉苹果酸酶活性与肌内脂肪含量呈显著正相关,与肉色呈正相关,而与滴水损失和剪切力呈显著负相关。     

Assessing hygiene proficiency on organic and conventional pig farms regarding pork safety: A pilot study in Finland

A key element in the EU Common Agricultural Policy (CAP) will be a single farm payment system that is linked to compliance with rules on, for instance hygiene standards. However, there are no recommended methods for assessing the hygiene proficiency of pig production farms. The present study was undertaken to develop a method for this purpose. A first implementation was done on pilot scale; with a set of both conventional and organic pig farms (N = 15). Fifty hygiene-related factors were selected, especially with reference to the possible proliferation of enteric pathogens Listeria monocytogenes, Yersinia enterocolitica and Yersinia pseudotuberculosis. The factors were allocated into 8 evaluation categories: (1) general production management, (2) animal density, (3) the outdoor area for pigs, (4) pest and pet animals, (5) general hygiene in the piggery, (6) pen hygiene, (7) feed production hygiene, and (8) feeding hygiene. A farm questionnaire and a supplementary on-site observation form were devised, and one arbitrary scale of hygiene points assigned for each factor. In addition to the mailed questionnaires, one person subsequently visited all the farms, perused the questionnaire with the owners and completed the observation form. The hygiene scores of the farms in each evaluation category were compared both without weighting and with weighting based on expert opinions. The method proved to be feasible and applicable to different types of production.     

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

A quantitative screening method to detect rater-introduced bias in clinical ratings

Background

We suggest a ‘screening test’ to examine large data files with clinical ratings for the occurrence of rater-introduced bias prior to using the data for quantitative analyses. The test is based on a statistical model in which a well-standardized interval-scale outcome (for example, milk yield) is related to clinical ratings (for example, body condition scores) obtained from multiple contexts (for example, dairy herds).

Findings

84,968 calvings from 279 herds, with subsequent body condition scores performed by 117 veterinarians within the first 21 days postpartum were analyzed with a multilevel random coefficient regression model. The model included an independent variable, where body condition score was centered within veterinarian. This is a so-called comparison effect to describe possible rater-introduced bias in the body condition scores. A highly significant comparison effect was found for second and older parities, indicating occurrence of possible rater-introduced bias in this large multi-herd data file.

Conclusions

A within-group centering technique (the comparison effect) appeared to be useful for discriminating between biased and unbiased clinical scores. In some cases, this test for bias should prevent further analysis of the data and divert the focus of study to the calibration of raters or alternative study designs.     

Comparison of bupivacaine femoral and sciatic nerve block versus bupivacaine and morphine epidural for stifle surgery in dogs

ObjectiveTo evaluate the efficacy of combined femoral and sciatic nerve blocks as an alternative to epidural anesthesia and analgesia in dogs undergoing stifle surgery under general anesthesia.Study designProspective, blinded, randomized, clinical comparison.AnimalsTwenty dogs weighing 37 ± 11 (mean ± SD) kg, aged 3 (1–8) [median (minimum–maximum)] years undergoing elective unilateral tibial-plateau leveling osteotomy.MethodsDogs were assigned randomly to receive either epidural anesthesia (bupivacaine 0.5%, 0.5 mg kg^?1 + morphine 0.1%, 0.1 mg kg^?1, in 0.2 mL kg^?1; EPID) or femoral and sciatic nerve blocks (Bupivacaine 0.5%, 0.1 mL kg^?1, was administered at each site; F + S) guided by electrolocation. All patients received a standard general anesthesia technique. Pain and sedation were scored (on scales of 0–10 and 0–3, respectively) pre-operatively, at extubation, and at 1, 4 and then every 4 hours thereafter up to 24 hours. Postoperatively, hydromorphone was administered to any patient with a pain score of >5 or whenever the blinded caregiver determined that more hydromorphone was necessary. Intraoperative heart rate (HR), mean arterial pressure (MAP), end tidal isoflurane (FE′ISO), body temperature, post-operative pain scores, time to first hydromorphone dose after surgery, time to first feeding, time to first drinking, time to first urination, time to first ambulation (walk on a lead) and cumulative dose of hydromorphone were recorded.ResultsIntra-operatively, FE′ISO and MAP were significantly lower in the EPID group (p = 0.05 and p = 0.04, respectively). Postoperatively, the cumulative hydromorphone consumption (p = 0.04) and the incidence of urinary retention (p = 0.03) were higher in the EPID group.Conclusion and clinical relevance F + S is a practical alternative to EPID that produces less urine retention and reduces opioid consumption in the 24 hours after surgery. EPID might be associated with a lower isoflurane requirement and lower systemic blood pressure.     

过渡期护理模式对重度颅脑损伤患者干预效果观察

目的 分析应用过渡期护理模式对重度颅脑损伤患者经ICU转入普通病房的效果,确保临床护理的精准性和延续性,以促进患者加快康复。方法 随机从我院2017年4月到2019年4月期间ICU的重度颅脑损伤患者中选取120名,分为实验组和对照组,对照组患者以常规性的护理干预模式,实验组则予以过渡期护理干预模式。观察两组患者发生不良事件的概率,SAS评分、重返ICU概率。结果 实验组患者发生不良事件的概率（3.33%）、SAS评分为（50.36±7.17）分、重返ICU概率（1.67%）均低于对照组,P<0.05;结论 过渡期护理干预模式对重度颅脑损伤患者有较为积极的作用,可减少不良事件的发生,减缓患者的焦虑情绪,促进患者康复。     

Correlations between acute radiation scores and pain scores in canine radiation patients with cancer of the forelimb

Objective To determine whether there is a correlation between skin acute radiation score (ARS) and pain scores and to determine if skin ARSs can be used to predict future pain scores and increased need for analgesia in dogs undergoing radiation therapy for cancer of the forelimb. Study design Prospective observational study. Animals Seven middle‐aged dogs of various breeds with cancer of the forelimb. Methods Each neoplasm was surgically removed and a histologic diagnosis was obtained. Curative intent radiation therapy was initiated 2½–4½ weeks after surgery. Curative intent radiation therapy was delivered as prescribed. Two trained observers scored the dogs using a visual analog pain scale (VAS), Glasgow composite measure of pain scale, short form (GCMPS) and skin ARS prior to each day’s therapy. Daily scores were averaged and scatter plots were developed. Generalized estimating equation regressions were used to calculate standard error, 95% confidence interval, and p‐values for each relationship. Confidence and prediction bands were plotted. Results A statistically significant correlation between skin ARS and VAS and GCMPS pain scores was identified indicating that as the skin ARS increased so did the pain scores. A general correlation between VAS and GCMPS scores was observed. Early (fraction days 1–6) GCMPS scores were significantly influenced by anxiety behavior unrelated to pain. Skin ARS was found to predict precisely current and future presence of pain, but could only predict a range of potential future pain scores based on the pain management approach in use during this study. Conclusions Skin ARS can provide valuable information for initiating preemptive analgesia and intensifying pain management during curative intent radiation therapy. Daily pain scoring with an acceptable pain scale should be used in conjunction with the skin ARS to improve patient pain management. Clinical relevance Pain is an anticipated consequence of curative intent radiation therapy. Understanding the correlation between pain and skin ARS may facilitate more effective pain management.